WO2021090910A1 - Molecular structure altering agent for detecting protein aggregates, detection method thereof, medical equipment cleaning agent, soil cleaning agent and soil cleaning method - Google Patents

Abstract

A molecular structure altering agent which interacts specifically with protein aggregates and detects said protein aggregates, and a detection method of protein aggregates are provided. Further, a cleaning agent and a cleaning method are provided for cleaning soil and medical equipment contaminated by protein aggregates. This molecular structure altering agent contains, as the compound that detects protein aggregates, a fluorine alcohol or a compound represented by general formula (1). (In general formula (1), a is 0 or 1; if a=1, then R¹ is a hydrogen atom and R² is a hydroxyl group, or R¹ is a hydroxyl group and R² is a hydrogen atom; if a=0, then R² is an oxygen atom that forms a divalent bond with a carbon atom, R³ is CH_lCl_mF_n with l, m and n integers from 1 to 3 and l+m+n=3, and R⁴ is CH_sCl_tF_u, with s, t and u integers from 1 to 3 and s+t+u=3.)

Description

Molecular structure altering agent for detecting protein aggregation, its detection method, cleaning agent for medical equipment, soil cleaning agent and soil cleaning method

The present invention relates to a molecular structure altering agent for detecting a protein that can be a pathogen of a disease group caused by a change in the higher-order structure of a protein such as prion disease, and a method for detecting the same. The present invention further relates to a molecular structure altering agent and a detection method for detecting a protein having a normal three-dimensional structure and a protein having an abnormal three-dimensional structure that can be a pathogen of a disease by a difference in sensitivity. Alternatively, the present invention relates to a cleaning agent for medical devices and a method for cleaning medical devices. Alternatively, the present invention relates to a soil cleaning agent and a method for cleaning soil.

Protein becomes a molecule that functions only when it forms the correct three-dimensional structure. However, if the correct three-dimensional structure is not formed, the protein causes fibrosis and aggregation to lose its original function or acquire new toxicity, thereby inhibiting various metabolic pathways in the living body. , Eventually can be fatal. These diseases are called protein misfolding diseases, and in the area of neurological diseases, prion disease, Alzheimer's disease, Parkinson's disease, Levy body dementia and the like are included. Cataracts, age-related macula, etc. are included in the field of ophthalmology. Amyloidosis is included in systemic organs.

Prion protein (PrP) is considered to be a pathogen that causes prion disease, which is a neurodegenerative disease. Prion's disease is a common veterinary infectious disease, with sporadic Creutzfeldt-Jakob disease (CJD), hereditary Gerstmann-Sträsler-Scheinker syndrome (GSS) and fatal familial insomnia (FFI) in humans. ), Infectious variant CJD (vCJD). Bovine spongiform encephalopathy (BSE) is known in cattle, scrapie in sheep, and deer chronic wasting disease (CWD) in deer. Prion disease also develops in cats, minks, and cheetahs. Protein conformational changes to normal type prion protein (PrP ^C) from the abnormality-type prion protein (PrP ^Sc), rather than immediately not cause prion infection, although there is debate still for these correlations, the development of prion disease It has been clarified that ^{PrP Sc is involved in.} However, no radical cure has been found to date. (Non-Patent Document 1, Non-Patent Document 2)

Protein misfolding disease has already been found to have fibrosis and aggregation formation derived from proteins with abnormal three-dimensional structure in the living body long before the first clinical symptoms appear. However, the pathophysiology of these diseases has progressed considerably when the person or family member notices the clinical symptoms, and early diagnosis is strongly required from the viewpoint of prevention of onset and treatment options. The current diagnostic criteria are, for example, sporadic prion disease, 1) showing progressive clinical symptoms, 2) showing cerebral atrophy on MRI imaging, and 3) 14- in cerebrospinal fluid. Criteria such as an increase in 3-3 protein and total tau protein are useful, but these diagnostic criteria are insufficient for early diagnosis, and further diagnostic methods are required. (Non-Patent Document 3)

Further, in industrial animals such as cattle and goats, a biopsy method that can eliminate individuals suffering from prion disease before sending them to a slaughterhouse is desired from the viewpoint of ensuring food safety. However, at present, there is no way to diagnose prion disease in vivo. Currently, the producer either senses the animal's anomaly or is found in a pre-slaughter test. In addition, even if it appears asymptomatic, it may be detected by surveillance of health and cattle. Furthermore, ^{although there are several antibodies that detect PrP Sc} , all of them can only be used for definitive diagnosis after death.

^{In prion disease, it is often considered that PrP Sc} accumulates as protein aggregates inside and outside nerve cells in the brain and exhibits pathogenicity, especially in the central nervous system in the living body. PrP ^Sc is has exactly the same amino acid sequence as PrP ^C, the three-dimensional structure of the protein are very different, PrP ^C is contains many α- helical structure, the ^{PrP Sc} contains many β- sheet structure It has been. Therefore, PrP ^Sc becomes a poorly soluble and poorly degradable protein and exhibits resistance to degradation by proteolytic enzymes such as proteinase K. Conformational conversion mechanism from PrP ^C to ^{PrP Sc} is not yet clear. In addition, it is said that changes in the three-dimensional structure have neurotoxicity and destroy nerve cells and the like over time, but the details are still unknown. Alzheimer's disease, Parkinson's disease, Levy body dementia, etc. are also caused by amyloid β protein, tau protein, α-synuclein, etc. as protein aggregates, respectively. Is believed to be caused by.

Unlike systemic organs, the central nervous system is composed of a group of cells that have completed differentiation, so once a group of proteins with three-dimensional structural abnormalities accumulates, they are not metabolized and remain in nerve cells.

Therefore, research and development of treatment methods and diagnostic methods for protein misfolding diseases is based on a strategy that focuses on persistent and persistent protein aggregates that accumulate in the body. In particular, in research and development intended for early diagnosis, the focus is on how to detect the formation of these protein aggregates and lead to prevention and early treatment of the onset. Patent Document 1 discloses a measurement method capable of detecting a 14-3-3 protein γ isoform in a biological sample with high sensitivity by an ELISA method (enzyme-linked immunosorbent assay) instead of the conventional Western blotting method. There is. The 14-3-3 protein γ isoform is a marker capable of detecting rapid nerve destruction in prion disease, and is known to be detected in cerebrospinal fluid in progressive neuropathy.

Patent Document 2 discloses a compound labeled with a radionuclide as a diagnostic imaging probe for prion protein-accumulating diseases. This allows imaging of prion proteins using positron emission tomography (PET). The problem with PET is that there are restrictions on the inspection sensitivity and the inspection time. Considering the half-life of the compound labeled with the radionuclide, the labeled compound must be synthesized by an automatic synthesizer immediately before the test. Patent Document 3 discloses an RNA aptamer, which is a nucleic acid molecule that binds to an abnormal prion-derived fibril. RNA aptamers bound to fibrils can be detected by PET or a fluorescence detector if they are labeled in advance, and even if they are not labeled, they can be detected by a crystal oscillator or a surface plasmon resonance detector.

Japanese Unexamined Patent Publication No. 2013-101047 Japanese Unexamined Patent Publication No. 2011-63574 Japanese Unexamined Patent Publication No. 2006-320289

Most of the conventional diagnostic methods for protein misfolding diseases are mainly based on clinical symptoms, followed by diagnostic imaging by MRI, and new diagnostic methods have been developed and proposed, but simpler and more sensitive examination methods. And diagnostic methods were needed. One embodiment of the present invention has been made in view of the above, and an object of the present invention is to provide a molecular structure altering agent capable of specifically interacting with a protein aggregate and detecting it, and a method for detecting the same. .. Alternatively, one embodiment of the present invention aims to provide a cleaning agent for cleaning a medical device contaminated with protein aggregates and a method for cleaning the medical device. Alternatively, one embodiment of the present invention aims to provide a cleaning agent for cleaning soil contaminated with protein aggregates and a method for cleaning soil.

As a result of diligent studies by the present inventors, when the fluoroalcohol 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) is brought into contact with the protein aggregate, the HFIP becomes the protein aggregate. It was found that it interacts specifically with and acts as a molecular structure altering agent to change the three-dimensional structure of protein aggregates.

The present invention includes the following aspects.

[1] A molecular structure altering agent containing a fluoroalcohol or a compound represented by the following general formula (1) as a compound for detecting protein aggregates.

(In the general formula (1), a is 0 or 1,
When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
Satisfy l + s <6. )

[2] Fluoro-based alcohols are represented by the general formula RfCH ₂ OH or RfRf'CHOH, Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms, and Rf and Rf'are different or the same. A molecular structure altering agent.

[3] A molecular structure altering agent in which the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.

[4] A molecular structure altering agent in which the concentration of a fluoroalcohol or a compound represented by the general formula (1) when acting on a protein aggregate is in the range of 10 pM to 100 mM.

[5] A molecular structure altering agent used for diagnosing protein misfolding disease.

[6] Protein misfolding disease includes sporadic Creutzfeldt-Jakob disease (CJD) in humans, hereditary Gerstmann-Sträsler-Scheinker syndrome (GSS), lethal familial insomnia (FFI), and medical doctors. Alzheimer, a prion disease in animals that can develop prion disease such as primary and dietary variants CJD (vCJD), bovine spongiform encephalopathy (BSE), sheep scrapy, deer chronic wasting disease (CWD), cats, minks, and cheetahs. Diseases, Parkinson's disease, Levy body dementia, neurodegenerative diseases included in the spectrum of frontotemporal lobar degeneration (FTLD), muscle atrophic lateral sclerosis, Huntington chorea, polyglutamine disease, cataracts, aging A molecular structure altering agent that is one disease selected from the group consisting of sex yellow spot and systemic amyloidosis.

[7] A method for detecting protein aggregates, which comprises allowing a fluoroalcohol or a compound represented by the following general formula (1) to act on the protein aggregates.

(In the general formula (1), a is 0 or 1,
When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
Satisfy l + s <6. )

[8] Fluoro-based alcohols are represented by the general formula RfCH ₂ OH or RfRf'CHOH, Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms, and Rf and Rf' are different or the same as each other. Method for detecting protein aggregates.

[9] A method for detecting protein aggregates in which the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.

[10] A method for detecting protein aggregates in which the concentration of a fluoroalcohol or a compound represented by the general formula (1) when acting on protein aggregates is in the range of 10 pM to 100 mM.

[11] A method for detecting protein aggregates used for diagnosing protein misfolding disease.

[12] Protein misfolding disease includes sporadic Creutzfeldt-Jakob disease (CJD) in humans, hereditary Gerstmann-Sträsler-Scheinker syndrome (GSS), lethal familial insomnia (FFI), and medical doctors. Alzheimer, a prion disease in animals that can develop prion disease such as primary and dietary mutant CJD (vCJD), bovine spongiform encephalopathy (BSE), sheep scrapy, deer chronic wasting disease (CWD), cats, minks, and cheetah Diseases, Parkinson's disease, Levy body dementia, neurodegenerative diseases included in the spectrum of frontotemporal lobar degeneration (FTLD), muscle atrophic lateral sclerosis, Huntington chorea, polyglutamine disease, cataracts, aging A method for detecting protein aggregates, which is one disease selected from the group consisting of sex yellow spot and systemic amyloidosis.

[13] A test kit for detecting protein aggregates containing a fluoroalcohol or a compound represented by the following general formula (1) as a compound for detecting protein aggregates.

(In the general formula (1), a is 0 or 1,
When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
Satisfy l + s <6. )

[14] The fluoroalcohol of the test kit is represented by the general formula RfCH ₂ OH or RfR f'CHOH.
Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms.
Rf and Rf'are different or the same as each other.

[15] The fluorine-based alcohol in the test kit is 1,1,1,3,3,3-hexafluoro-2-propanol.

[16] The concentration of the fluoroalcohol or the compound represented by the general formula (1) when acting on the protein aggregate of the test kit is in the range of 10 pM to 100 mM.

[17] An inspection device for detecting protein aggregates, which comprises a detection unit containing a fluoroalcohol or a compound represented by the following general formula (1) as a compound for detecting protein aggregates.

(In the general formula (1), a is 0 or 1,
When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
Satisfy l + s <6. )

[18] The fluorinated alcohol of the inspection device is represented by the general formula RfCH ₂ OH or RfR f'CHOH.
Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms.
Rf and Rf'are different or the same as each other.

[19] The fluoroalcohol of the inspection device is 1,1,1,3,3,3-hexafluoro-2-propanol.

[20] The concentration of the fluorinated alcohol or the compound represented by the general formula (1) when acting on the protein aggregate of the inspection device is in the range of 10 pM to 100 mM.

[21] A cleaning agent for medical instruments containing a fluorinated alcohol or a compound represented by the following general formula (1).

(In the general formula (1), a is 0 or 1,
When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
Satisfy l + s <6. )

[22] In the cleaning agent for medical instruments, the fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH, and Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms. Rf'is different or the same as each other.

[23] In the cleaning agent for medical instruments, the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.

[24] A soil cleaning agent containing a fluorinated alcohol or a compound represented by the following general formula (1).

(In the general formula (1), a is 0 or 1,
When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
Satisfy l + s <6. )

[25] In the soil cleaning agent, the fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH, Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms, and Rf and Rf' Are different or the same as each other.

[26] In the soil cleaning agent, the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.

[27] A method for cleaning soil using the soil cleaning agent.

According to one embodiment of the present invention, a molecular structure altering agent that alters a protein three-dimensional structure with respect to protein aggregates accumulated in humans and animals is provided, protein aggregates are detected, and protein misfolding disease is diagnosed. Can be used. Alternatively, according to one embodiment of the present invention, it is possible to provide a cleaning agent for cleaning a medical device contaminated with protein aggregates, and to provide a method for cleaning the medical device using the cleaning agent. Alternatively, according to one embodiment of the present invention, it is possible to provide a cleaning agent for cleaning soil contaminated with protein aggregates, and to provide a method for cleaning soil using the cleaning agent.

It is a schematic diagram which shows the cleaning apparatus 1 which concerns on one Embodiment of this invention. In cell cultures from murine neuroblastoma, a photograph of Neuro 2a (N2a) cells and ScNeuro 2a (ScN2a) cell culture plate was tested sensitivity at each concentration of HFIP for cells producing PrP ^Sc producing a PrP ^C is there. This is the result of Western blotting evaluation of the resistance of N2a cells and ScN2a cells to proteinase K after cell culture under the condition of adding or not adding HFIP. It is a schematic diagram which shows the inspection apparatus 100 which concerns on one Embodiment of this invention.

The compound that alters the molecular structure of human and animal protein aggregates of the present invention is a fluoroalcohol or a compound represented by the following general formula (1).

In the general formula (1), when a is 0 or 1 and a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group. Alternatively, R ¹ is a hydroxyl group and R ² is a hydrogen atom. When a = 0, R ² is an oxygen atom that forms a double bond with a carbon atom. R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3. Further, R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, and s + t + u = 3 is satisfied. It should be noted that l + s <6 is satisfied.

In one embodiment, the molecular structure altering agent contains a fluorinated alcohol or a compound represented by the above general formula as an active ingredient. In the present specification, the "molecular structure modifier" specifically acts on a protein containing a large amount of β-sheet structure to unfold the molecular structure from the β-sheet structure to form an α-helix structure. It contains an inducing compound and can be suitably used for detection of protein aggregates and simple screening and diagnosis of protein misfolding diseases including neurodegenerative diseases. In one embodiment, the molecular structure altering agent contains two or more selected from a fluoroalcohol and a compound represented by the above general formula.

In one embodiment, the compound represented by the general formula (1) may be selected from the compound group represented by the following compounds 1 to 3.
[Compound group]

The fluoroalcohol of the present invention is represented by the following general formula (2) or (3).

RfCH ₂ OH ・ ・ ・ (2)
RfRf'CHOH ・ ・ ・ (3)

In the above general formula, Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms.

Among the compounds represented by the general formula (2) or (3), 1,1,1,3,3,3-hexafluoro-2-propanol (hereinafter, may be referred to as HFIP), 2,2. , 2-Trifluoroethanol, 2,2,3,3,3-pentafluoro-1-propanol can be exemplified. In particular, HFIP is suitable as a compound that alters the molecular structure of protein aggregates. In one embodiment, the molecular structure altering agent may contain at least one of the compounds represented by the general formulas (1), (2) or (3) and a solvent.

Examples of the solvent capable of diluting the compound represented by the general formula (1), (2) or (3) include water, physiological saline, Ringer's solution, phosphate buffered saline (PBS), methanol, and the like. Examples include, but are not limited to, ethanol, isopropanol, acetone, toluene, dimethyl sulfoxide and the like. In one embodiment, the molecular structure altering agent contains the compound represented by the general formula (1), (2) or (3) and the above solvent. HFIP is a colorless and transparent liquid with a melting point of -3.3 ° C and a boiling point of 58.6 ° C, and is soluble in most solvents. Therefore, HFIP should be adjusted to an arbitrary concentration by diluting with a solvent. Can be done.

The concentration of the fluoroalcohol or the compound represented by the general formula (1) when acting on the protein aggregate is preferably in the range of 10 pM or more and 100 mM or less. The time for allowing the fluoroalcohol or the compound represented by the general formula (1) to act on the protein aggregate is not particularly limited.

In one embodiment, it is represented by a fluorinated alcohol or a compound represented by the general formula (1), or instead of the fluorinated alcohol or the compound represented by the general formula (1). A compound in which the hydroxy group in the compound is protected by a protecting group may be used. This protecting group suppresses the decomposition of the fluoroalcohol or the compound represented by the general formula (1) in the process of acting on the protein aggregate, and this compound is the fluoroalcohol or the general when acting on the protein aggregate. It suffices if it is converted into the compound represented by the formula (1). Examples of such protecting groups include functional groups generally used as protecting groups for hydroxy groups (see, for example, "Protecting Group Chemistry" by Jeremy Robertson, published by Oxford University Press), sugar chains, peptides and the like. Examples of the functional group used as the protective group include acetal-based functional groups such as an alkoxyalkyl group (for example, methoxymethyl group and ethoxyethyl group) and 2-tetrahydropyranyl group, and an alkanoyl group (for example, acetyl group) and an aloyl group. Acrylic functional group such as (for example, arylmethyl group such as benzyl group, benzoyl group), silyl ether functional group such as alkylsilyl group (for example, trimethylsilyl group, triethylsilyl group, tert-butyldimethylsilyl group) However, the present invention is not limited thereto.

[Method for detecting protein aggregation]
The molecular structure altering agent containing the fluoroalcohol of the present invention or the compound represented by the general formula (1) can be used for diagnosing protein misfolding diseases, particularly neurodegenerative diseases, and at a pharmaceutically acceptable concentration. It can be contained as an active ingredient to form a test kit. Here, the pharmaceutically acceptable concentration is the same as the concentration that alters the molecular structure of the protein aggregate, and specifically, the concentration is preferably in the range of 10 pM or more and 100 mM. In one embodiment, the test kit comprises HFIP and the solvent described above.

As used herein, protein misfolding diseases in the nervous system include sporadic Creutzfeldt-Jakob disease (CJD), hereditary Gerstmann-Sträsler-Scheinker syndrome (GSS), and lethal families in prion disease. Onset of prion diseases such as sexual insomnia (FFI), iatrogenic and dietary mutant CJD (vCJD), bovine spongiform encephalopathy (BSE), sheep scrapy, deer chronic wasting disease (CWD), cats, minks, cheetahs, etc. Possible animal prion diseases can be exemplified. Others include Alzheimer's disease, Parkinson's disease, Lewy body dementias, neurodegenerative diseases in the frontotemporal lobar degeneration (FTLD) spectrum, muscle atrophic lateral sclerosis, Huntington's chorea, and polyglutamine disease. Can be exemplified. In the field of ophthalmology, cataracts, age-related macula and the like can be exemplified. Amyloidosis is included in systemic organs.

In neurodegenerative diseases, proteins showing a normal three-dimensional structure cause higher-order structural changes in proteins with an abnormal three-dimensional structure, which accumulate inside and outside nerve cells in the brain as protein aggregates, especially in the central nervous system. It is believed to be pathogenic. In sporadic and infectious prion diseases, the amino acid sequence of a protein does not change during this process, only its conformation. Normal proteins contain a large amount of α-helix structure as a secondary structure, but proteins with an abnormal three-dimensional structure contain a large amount of β-sheet structure, which makes them sparingly soluble and persistently degradable, such as proteinase K. Shows resistance to proteolytic enzymes.

The fluoroalcohol contained in the molecular structure altering agent of the present invention, especially HFIP, acts specifically on the β-sheet structure of a protein to unfold the molecular structure from the β-sheet structure and induce it into an α-helix structure. However, in neurodegenerative diseases, it can be an effective molecular structure modifier for protein aggregates accumulated in the living body, especially in the central nervous tissue. In addition, HFIP is a low molecular weight compound having a molecular weight of 168, and has a molecular size capable of crossing the cerebral blood barrier that controls substance exchange between blood and brain tissue fluid (generally, a molecule having a molecular weight of more than 500). (It is said that it cannot pass through), and it is thought that it moves into the brain and functions as a molecular structure modifier.

In one embodiment, biological samples that can be used in the method for detecting protein aggregates of the present invention include organs, biological tissues, and cells suspected of being infected with protein misfolding disease, such as the brain, spinal cord, and the like. Tonsillar, nerve junction, spleen, heart, liver, lung, eyeball, placenta, testis, lymphatic tissue, muscle tissue, etc., or nerve cells, blood cells, muscle cells, and blood (including liquid components of plasma and serum), Body fluids such as cerebrospinal fluid (medullary fluid) and urine are not particularly limited as long as protein aggregates can be detected by the method for detecting protein aggregates of the present invention. Blood, cerebrospinal fluid, and the like are preferable because they are relatively minimally invasive samples. In addition, tonsils, nerve junctions, etc. are tissues that are easier to collect than the brain, and are suitable for diagnosis of industrial animals.

Biological samples infected with protein misfolding disease include cells containing protein aggregates, and by utilizing the difference in susceptibility between these cells and a fluoroalcohol or a compound represented by the general formula (1), It can be used for simple screening and diagnosis of protein misfolding disease. In one embodiment, for example, cells suffering from protein misfolding disease lead to cell death depending on the concentration of fluoroalcohol or the compound represented by the general formula (1) as compared to normal cells. In order to increase the detection sensitivity of protein aggregates, biological samples collected from living tissues may be used after culturing in vitro. The suitable biological sample differs depending on the protein to be detected, and is not particularly limited. For example, PrP ^Sc is said to be present in cerebrospinal fluid, and amyloid beta such as Alzheimer's disease is also present in blood.

The detection of protein aggregates of the present invention can be used not only for simple screening and diagnosis of protein misfolding disease, but also for tests for preventing infection by blood for transfusion, blood products, spinal fluid, organs for transplantation, and the like. Currently, the blood donors have restrictions on the tests for prion infection of blood for transfusion, such as the history of overseas stays and disease history of blood donors.

The test for altering the molecular structure of a protein aggregate using the fluoroalcohol of the present invention or the compound represented by the general formula (1) is a method capable of evaluating a protein having a normal molecular structure and a protein having an abnormal molecular structure, respectively. If there is, there is no particular limitation.

In one embodiment, when the above tissue or cell is used as a biological sample, protein aggregates are accumulated due to the difference in sensitivity between normal cells and cells in which protein aggregates are accumulated to the molecular structure altering agent of the present invention. The presence or absence of cells can be detected. For example, the presence or absence of cells in which protein aggregates have accumulated can be detected by allowing cells to act on the molecular structure altering agent of the present invention for a predetermined time and then evaluating live cells or dead cells by cell staining.

In one embodiment, when the above tissue or cell is used as a biological sample, the protein aggregate is due to the difference in sensitivity of the protein having a normal structure to the molecular structure altering agent of the present invention and the protein aggregate to a proteolytic enzyme. The presence or absence of accumulated tissue or cells can be detected. For example, when the cell lysate is treated with a proteolytic enzyme, the protein aggregate is not decomposed by the proteolytic enzyme, and a band derived from the protein aggregate is detected by electrophoresis. On the other hand, when the cell lysate is treated with the molecular structure alteration agent of the present invention and then treated with a proteolytic enzyme, the protein aggregates altered by the molecular structure alteration agent are decomposed by the proteolytic enzyme, and a band of the protein aggregates is electrophoresed. Dilutes or disappears. Therefore, the presence or absence of tissues or cells in which protein aggregates have accumulated can be detected by the difference in sensitivity to proteolytic enzymes before and after treatment with the molecular structure altering agent of the present invention. Further, even when the protein on which the protein aggregation is formed is unknown, the protein can be identified by, for example, mass spectrometry by a shotgun method or a combination of two-dimensional electrophoresis and mass spectrometry. ..

[Inspection device]
In one embodiment, it is possible to provide an inspection device containing the above-mentioned fluorinated alcohol according to the present invention or the compound represented by the general formula (1) in the detection unit.

FIG. 4 is a schematic view showing an inspection device 100 according to an embodiment of the present invention. The inspection device 100 includes, but is not limited to, for example, an input unit 110, a detection unit 120, a storage unit 130, a display unit 140, a control unit 150, a calculation unit 160, a power supply device 170, and a communication unit 180. .. Further, the inspection device 100 may be connected to the server 300 via the network 200.

The input unit 110 is, for example, a device for inputting information such as an inspection target and inspection conditions into the inspection device 100, and may be composed of a known input device such as a keyboard, mouse, or touch panel. The information to be tested may be, for example, the above-mentioned cell or tissue name, patient name, ID, or the like. Further, the test conditions may be conditions related to the test such as the name and concentration of the fluoroalcohol or the compound represented by the general formula (1), the date and time of the test, and the number of cells according to the present invention.

The detection unit 120 is a device for detecting the state of cells or protein aggregates by adding, for example, the fluoroalcohol according to the present invention or the compound represented by the general formula (1) to cells to be inspected. In one embodiment, the detection unit 120 may contain a fluorinated alcohol according to the present invention or a compound represented by the general formula (1). For example, the detection unit 120 supplies a fluorine-based alcohol according to the present invention or a compound represented by the general formula (1) to cells or the like placed in a container for detection to detect the state of cells or protein aggregates. You may. In the present specification, the detection unit 120 supplies a fluoroalcohol or a compound represented by the general formula (1) according to the present invention to perform a series of processes for detecting the state of cells and protein aggregates. The means of the above may be shown, and the detection unit 120 may detect the state of cells or protein aggregates by supplying the fluoroalcohol according to the present invention or the compound represented by the general formula (1) by a person. In addition, the detection unit 120 may include an electrophoresis device for detecting the state of protein aggregates. The detection unit 120 may be provided with an imaging device in order to detect the state of cells and protein aggregates. As the image pickup apparatus, for example, a known CCD image sensor or CMOS image sensor can be used, and therefore detailed description thereof will be omitted.

The storage unit 130 includes a main storage device and an auxiliary storage device. Since a known memory can be used as the main storage device, detailed description thereof will be omitted. Further, since a known hard disk, solid state drive (SSD), or the like can be used as the auxiliary storage device, detailed description thereof will be omitted. The storage unit 130 can store data such as an operating system, application software, and detection results for the inspection device 100. The auxiliary storage device is not essential. For example, when the inspection device 100 is connected to the server 300 via the network 200, data such as application software and inspection results may be stored in the server 300.

Since the display unit 140 is a device for displaying information for operating the inspection device 100, inspection results, and the like, and a known display can be used, detailed description thereof will be omitted.

The control unit 150 is composed of a central processing unit and a program for controlling the inspection device 100. The control unit 150 includes, for example, an operating system, application software, or modules.

The calculation unit 160 executes a calculation based on the information of the inspection target input from the input unit 110, the inspection conditions, and the state of the cells and protein aggregates acquired by the detection unit 120, and provides the inspection result. The calculation unit 160 includes application software or a module used for calculation.

Since the power supply device 170 is a device for supplying power to the inspection device 100 from the outside and a known power supply device or battery can be used, detailed description thereof will be omitted.

The communication unit 180 is a device for the inspection device 100 to communicate with an external device by wire or wirelessly. The communication unit 180 can be connected to the server 300 or another device (not shown) via a known local area network (LAN) or a network 200 such as the Internet.

In one embodiment, the inspection device 100 detects the state of cells to be inspected and the state of protein aggregates after the action of the fluorinated alcohol according to the present invention or the compound represented by the general formula (1). The risk of developing protein misfolding disease and the degree of progression of the pathological condition may be evaluated by detecting with the above and comparing with the control data or the standard value stored in the storage unit 130 or the server 300.

In one embodiment, the inspection device 100 compares the inspection result at the time of inspection (first time point) with the previous inspection result stored in the storage unit 130 or the server 300 (second time point), and compares the protein. The risk of developing misfolding disease and the degree of progression of the condition may be assessed.

In one embodiment, the inspection device 100 causes the neural network to perform machine learning processing using the teacher data stored in the storage unit 130 or the server 300, and uses the trained neural network to perform the inspection detected by the detection unit 120. Based on the subject's data, the risk of developing protein misfolding disease and the degree of progression of the condition may be evaluated. Here, the teacher data includes predetermined input data (detection results, etc.) related to each sample collected from a plurality of samples, and output data indicating that the organism from which the sample was collected may have protein misfolding disease. Including.

In one embodiment, the inspection device 100 may output the evaluation result by a printer via the communication unit 180, and the inspection device 100 may output the evaluation result from the communication unit 180 to the terminal 400 for an electronic medical record or a medical worker via the network 200. The evaluation result may be sent.

[Cleaning agent for medical equipment]
The molecular structure altering agent according to the present invention described above can be used as a cleaning agent for medical instruments in one embodiment. For example, the risk of CJD infection via instruments used in neurosurgery in CJD patients has been pointed out. The UK CJD Incident Panel ^{reports that the pathogenicity of PrP Sc} adhering to surgical instruments is removed by about 10 normal disinfections when normal disinfection is used. Therefore, it is difficult to sufficiently reduce the risk of CJD infection by the usual cleaning method of medical devices.

In one embodiment, the cleaning agent for medical instruments contains the above-mentioned molecular structure altering agent according to the present invention, and may further contain a solvent. The solvent can be selected from materials that can dilute the molecular structure altering agent, such as water, physiological saline, borate buffer, phosphate buffer (for example, phosphate buffered saline (also referred to as PBS)), acetate buffer, etc. Examples thereof include Tris buffer, HEPES buffer, methanol, ethanol, isopropanol, acetone, toluene, dimethyl sulfoxide, ethylene glycol, diethylene glycol and propylene glycol. These may be one or more, but are not limited thereto.

For example, HFIP is a colorless and transparent liquid with a melting point of -3.3 ° C and a boiling point of 58.6 ° C, and is soluble in most solvents. Therefore, HFIP can be adjusted to an arbitrary concentration by diluting with a solvent. can do. In one embodiment, the content (% by mass) of the solvent contained in the cleaning agent for medical instruments is 0% by mass or more and 99.9% by mass or less with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. It may be 0% by mass or more and 99% by mass or less, 0% by mass or more and 92% by mass or less is particularly preferable, and 0% by mass or more and 71% by mass or less is further preferable.

In one embodiment, the cleaning agent for medical instruments may contain additives in addition to the molecular structure altering agent and the solvent. Additives that can be added to medical device cleaning agents include surfactants, enzymes, chelating agents, enzyme stabilizers, blood coagulation inhibitors, metal corrosion inhibitors, low molecular weight polyols, cleaning aids (builders), and defoamers. Agents, pH regulators, fragrances, colorants, antioxidants, preservatives, bleaching agents, bleaching activators, corrosion inhibitors dispersants, thickeners, viscosity regulators, etc., but are limited to these. It's not a thing. The cleaning agent for medical instruments may contain one or more additives. In one embodiment, the cleaning agent for medical instruments can further improve the cleaning power by containing a molecular structure altering agent, a solvent, and the above-mentioned additive.

Surfactants (A) include nonionic surfactants (A-1), anionic surfactants (A-2), cationic surfactants (A-3), and amphoteric surfactants (A-). 4) and biosurfactant (A-5) are included.

Examples of the nonionic surfactant (A-1) include an alkylene oxide-added nonionic surfactant (A-1-1) and a polyhydric alcohol-type nonionic surfactant (A-1-2). Can be mentioned.

As the alkylene oxide adduct nonionic surfactant (A-1-1), a higher alcohol (8 to 18 carbon atoms) alkylene (2 to 4 carbon atoms, preferably 2) oxide adduct (1 active hydrogen) Per addition mole number 1 to 30), alkyl (carbon number 1 to 12) phenol ethylene oxide (hereinafter, ethylene oxide may be referred to as EO) adduct (addition mole number 1 to 30), higher amine (carbon) Numbers 8 to 22) alkylene (2 to 4 carbon atoms, preferably 2) oxide adduct (1 to 40 moles added per active hydrogen), fatty acid (8 to 18 carbon atoms) EO adduct (active hydrogen) 1 to 60 moles added per piece), polypropylene glycol (200 to 4000 molecular weight) EO adduct (1 to 50 moles added per active hydrogen), polyoxyethylene (3 to 30 repeating units) alkyl (6 to 20 carbon atoms) Allyl ether and sorbitan monolaurate EO adduct (1 to 30 moles added per active hydrogen) and sorbitan monooleate EO adduct (1 to 30 moles added per active hydrogen) 30) and other polyhydric (2-8 valent or higher) alcohols (2-30 carbon atoms) fatty acids (8-24 carbon atoms) ester EO adducts (1-30 adducts per active hydrogen) And so on.

Examples of the polyhydric alcohol-type nonionic surfactant (A-1-2) include polyvalent (2 to 8-valent or higher) such as glycerin monostearate, glycerin monooleate, sorbitan monolaurate and sorbitan monoolate. Examples thereof include fatty acid (8 to 24 carbon atoms) esters of alcohols (2 to 30 carbon atoms) and fatty acid alkanolamides such as lauric acid monoethanolamide and lauric acid diethanolamide.

As the anionic surfactant (A-2), an alkyl ether carboxylic acid having 8 to 24 carbon atoms or a salt thereof and an alkyl (poly) oxyethylene ether carboxylic acid having 8 to 24 carbon atoms or a salt thereof [(poly) oxy Ethylene (degree of polymerization = 1-100) sodium lauryl ether acetate and (poly) oxyethylene (degree of polymerization = 1-100) sodium lauryl sulfosuccinate, etc.], alkyl sulfate ester salt having 8 to 24 carbon atoms and 8 to 8 carbon atoms Twenty-four alkyl (poly) oxyethylene sulfate ester salts [sodium lauryl sulfate, lauryl (poly) oxyethylene (polymerization degree = 1 to 100) sodium sulfate and lauryl (poly) oxyethylene (polymerization degree = 1 to 100) sulfate-tri Ethanolamine salt, etc.], palm oil fatty acid monoethanolamide sodium sulfate sulfonate, alkylphenyl sulfonate with 8 to 24 carbon atoms [sodium dodecylbenzene sulfonate, etc.], alkylphosphate ester salt with 8 to 24 carbon atoms and carbon Alkyl (poly) oxyethylene phosphate ester salts of number 8 to 24 [sodium lauryl phosphate and (poly) oxyethylene (polymerization degree = 1 to 100) sodium lauryl ether phosphate, etc.], fatty acid salts [sodium lauryl sulfate and laurin Acid triethanolamine, etc.], acylated amino acid salt [sodium coconut oil fatty acid methyl taurine, coconut oil fatty acid sarcosin sodium, coconut oil fatty acid sarcosin triethanolamine, N-palm oil fatty acid acyl-L-glutamate triethanolamine, N -Palm oil fatty acid acyl-L-sodium glutamate, sodium lauroylmethyl-β-alanine, etc.].

Examples of the cationic surfactant (A-3) include quaternary ammonium salt type [stearyltrimethylammonium chloride, behenyltrimethylammonium chloride, distearyldimethylammonium chloride and lanolin fatty acid ethyl sulfate aminopropylethyldimethylammonium, etc.] and amine salts. Types [diethylaminoethylamide stearate, dilaurylamine hydrochloride, oleylamine, etc.] and the like can be mentioned.

Examples of the amphoteric tenside agent (A-4) include betaine-type amphoteric tenside agents [coconut oil fatty acid amidopropyldimethylaminoacetic acid betaine, lauryldimethylaminoacetic acid betaine, 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazole. Nium betaine, lauryl hydroxysulfobetaine, lauroylamide ethyl hydroxyethyl carboxymethyl betaine sodium hydroxypropyl phosphate, etc.], amino acid amphoteric tenside [β-lauryl aminopropionate, etc.] can be mentioned.

Examples of biosurfactant (A-5) include surfactin, ramnolipid, and salts thereof. Examples of the salt include alkali metal salts, alkaline earth metal salts, onium salts and the like.

As the surfactant (A), one type or two or more types can be used. When two or more kinds are used, the combination includes, for example, a nonionic surfactant (A-1) and an anionic surfactant (A-2), a nonionic surfactant (A-1) and a cation. Examples thereof include a combination of an amphoteric surfactant (A-3), a nonionic surfactant (A-1) and an amphoteric surfactant (A-4).

As the surfactant (A), from the viewpoint of detergency, the nonionic surfactant (A-1) can be used alone, and the nonionic surfactant (A-1) and the anionic surfactant (A-1) can be used alone. It is preferable to use it in combination with -2).

As the nonionic surfactant (A-1), an aliphatic alcohol (8 to 24 carbon atoms) ethylene oxide adduct (polymerization degree = 1 to 100) is preferable, and more preferably an aliphatic alcohol, from the viewpoint of detergency. Alcohol (12-18 carbon atoms) ethylene oxide adduct (polymerization degree 4-20), then more preferably aliphatic alcohol (12-15 carbon atoms) ethylene oxide adduct (polymerization degree = 8-12), particularly preferably. Is an 11 mol adduct of lauryl alcohol ethylene oxide.

The anionic surfactant (A-2) includes an alkylphenyl sulfonate having 8 to 24 carbon atoms, a fatty acid salt, an alkyl sulfate ester salt having 8 to 24 carbon atoms, and an alkyl sulfate ester salt having 8 to 24 carbon atoms from the viewpoint of detergency. Alkyl (poly) oxyethylene sulfate ester salt of, more preferably an alkylphenyl sulfonate having 12 to 16 carbon atoms and a fatty acid salt having 8 to 16 carbon atoms, and further preferably dodecylbenzenesulfonic acid monoethanol. Amine salt and sodium laurate.

The content (mass%) of the surfactant (A) contained in the cleaning agent for medical instruments is preferably 0% by mass or more and 10% by mass or less with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. , More preferably 0.1% by mass or more and 5% by mass or less.

Enzyme (B) includes protease (B-1), amylase (B-2), lipase (B-3) and cellulase (B-4), aminopeptidase and the like.

In the present specification, the protease (B-1) is an enzyme that catalyzes hydrolysis using a peptide or protein as a substrate. The protease (B-1) includes those of animal, plant or microbial origin, and those of microbial origin are preferable from the viewpoint of availability. Also included are chemically or genetically modified variants. The protease (B-1) may be a protease (alkaline protease) having an optimum pH on the neutral to alkaline side, and a plurality of proteases satisfying this condition can be used in combination. Examples of the protease (B-1) include serine protease (B-1-1), aspartic protease (B-1-2), cysteine protease (B-1--3) and metalloprotease (B-1--4). included.

Serine protease (B-1-1) is a protease having a serine residue as a catalytic residue, and is a chymotrypsin, trypsin, thrombin, plasmin, elastase, subtilisin (subtilisin E, subtilisin BPN'), kexin, and actinomycete (subtilisin E, subtilisin BPN'). It includes proteases derived from (streptomyces), proteases derived from subtilisin (Bacillus), proteases derived from filamentous fungi (Aspergillus), and the like. Specific examples thereof include trypsin derived from porcine pancreas, subtilisin Novo derived from Bacillus, subtilisin Carlsberg, subtilisin 309, subtilisin 147 and subtilisin 168. Serine protease (B-1-1) is a protease in which a serine residue is involved in the active center, and is inactivated by a drug such as diisopropylfluorophosphoric acid or phenylmethanesulfonylfloride that specifically binds to the serine residue. It has been known. Serine protease (B-1-1) does not require a reducing agent, is not affected by a metal chelating agent, and has an optimum pH of enzyme activity near neutrality, and is therefore preferably used in this embodiment. Be done.

Commercially available serine proteases (B-1-1) include Alcalase, Sabinase, Evalase, Esperase, Cannase, Ovozyme, Subtilisin A, PEM, PTN, Primase, Durazym manufactured by Novozymes, and Bioplase manufactured by Nagase Biochemical Industry Co., Ltd. , Protease N "Amano" manufactured by Amano Pharmaceutical Co., Ltd., Protease P "Amano", Actinase AS manufactured by Kaken Pharmaceutical Co., Ltd., KAP manufactured by Kao Co., Ltd. Examples thereof include pronase, TrypLE Select manufactured by Invitrogen Co., Ltd., and Accutase manufactured by Chemicon International. Further, the protease described in JP-A-2007-61101 can also be preferably used.

Aspartic protease (B-1-2) is a protease in which aspartic acid is present in the active center, and includes pepsin, cathepsin D, cathepsin E, renin, chymosin, and the like. Specific examples include pepsin derived from the human stomach. Aspartic protease (B-1-2) is a protease generally also called an acidic protease and has enzymatic activity in the acidic region. HFIP is suitable because it is acidic.

Cysteine protease (B-1--3) is a protease in which a thiol group is present in the active center, and includes papain, bromelain, ficin, actinidin, cathepsin B, cathepsin H, cathepsin L, caspase, ginger protease and the like. Since the active center of cysteine protease (B-1--3) is a thiol group, it is preferable to use a reducing agent such as cysteine or thiourea in combination. From the viewpoint of preventing oxidation by oxygen in the air, such a reducing agent is preferably added to the cleaning agent immediately before or during cleaning.

The metalloprotease (B-1--4) is a protease containing a metal ion in the active center, and examples thereof include thermolysin, matrix metalloproteinase, carboxypeptidase A, carboxypeptidase B, dispase, and collagenase. Examples of commercially available metalloproteases (B-1-4) include dispase manufactured by Worthington Biochemical Corporation. In one embodiment, when the metalloproteinase (B-1-4) and the chelating agent (C) described later are used in combination, it is preferable to use a metal-free chelating agent from the viewpoint of maintaining the activity of the metalloproteinase.

Among the above proteases (B-1), serine protease (B-1-1) and aspartic protease (B-1-2) are preferable, and subtilisin and plasmin are more preferable, from the viewpoint of long-lasting effect and detergency. Is. Of these, subtilisins derived from Bacillus Hallodurans and Bacillus clausii are preferable.

In one embodiment, the cleaning agent for medical instruments can clean the adhered protein stains more efficiently by containing the protease (B-1). The protease (B-1) may be contained in the cleaning agent for medical instruments, but the cleaning agent containing the protease (B-1) may be used in combination with the cleaning agent for medical instruments according to the present embodiment. From the viewpoint of enzyme stability, it is preferable to separately prepare a cleaning agent containing a protease (B-1) and use it in combination immediately before or during washing.

Amylase (B-2) includes those of bacterial or fungal origin. Also included are chemically or genetically modified variants. Examples of amylase (B-2) are described in detail in British Patent No. 1,296,839. Examples thereof include α-amylase obtained from a special strain of B. licheniformis. Examples of commercially available amylase (B-2) include Duramyl, Termamyl, Fungamyl and BAN manufactured by Novozymes, and Rapidase and Maxamyl P manufactured by Gist-Brocades.

Lipase (B-3) includes those of bacterial or fungal origin. Also included are chemically or genetically modified variants. Examples of lipases include Humicola langinosa lipase (European Patent No. 258 068 and European Patent No. 30 216), Rhizomucor miehei lipase and Candida (Candida). Patent No. 238, 023), C.I. C. ntarctica lipases A and B, Pseudomonas lipases (European Patent No. 214, 761), P. et al. P. pseudoalcaligenes and P. pseudoalcaligenes. Alcaligenes lipase (European Patent No. 218, 272), P. et al. P. cepasia lipase (European Patent No. 331, 376), P. et al. P. stutzeri lipase, P. stutzeri. P. Fluorescence lipase and Bacillus lipase (UK Pat. No. 1,372,034), B.I. B. subtilis lipase (Dartois et al. (1993), Biochemica et Biophysica Acta1131,253-260), B.I. B. stearothermophilus lipase (Japanese Patent Publication No. 64-744922) and B. B. pumilus lipase (International Publication No. 91/16422) can be mentioned.

Examples of commercially available lipase (B-3) include M1 Lipase, Luma fast and Lipomax from Genecore, Lipase and Lipase Ultra from Novozymes, and Lipase P "Amano" from Amano Enzyme.

Cellulase (B-4) includes those of bacterial or fungal origin. Also included are chemically or genetically modified variants. Cellulases include those disclosed in US Pat. No. 4,435,307 as fungal cellulases produced from Humicola insolence.

Examples of commercially available cellulases include Cellulase of Novozymes Co., Ltd. and KAC-500 (B) of Kao Corporation produced by the strain of Humicola insolence.

Of the above enzymes (B), protease (B-1) is preferable from the viewpoint of detergency.

In one embodiment, the enzyme (B) contained in the cleaning agent for medical instruments can contain two or more types. Examples of the combination containing two or more kinds include a combination containing two or more kinds of protease, protease and amylase, protease and lipase, or protease and amylase and lipase.

In one embodiment, the content (% by mass) of the enzyme (B) contained in the cleaning agent for medical instruments is 0% by mass or more and 10% by mass with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. The following is preferable, more preferably 0.05% by mass or more and 5% by mass or less, and particularly preferably 0.1% by mass or more and 3% by mass or less.

As the chelating agent (C), any of aminocarboxylic acid type, organic acid type, phosphonic acid type, phosphoric acid type and polycarboxylic acid type can be used. For example, aminopolyacetic acids such as nitrilotriacetic acid, iminodiacetic acid, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid, glycol etherdiaminetetraacetic acid, hydroxyethyliminodiacetic acid, triethylenetetraaminehexacetic acid, diencoric acid, etc. Organic acids such as salts, diglycolic acid, oxydisuccinic acid, carboxymethyloxysuccinic acid, citric acid, lactic acid, tartrate acid, oxalic acid, malic acid, gluconic acid, carboxymethyl succinic acid, carboxymethyl tartrate acid, glutamate diacetic acid, etc. Salt, aminotri (methylenephosphonic acid), 1-hydroxyethylidene-1,1-diphosphonic acid, ethylenediaminetetra (methylenephosphonic acid), diethylenetriaminepenta (methylenephosphonic acid) and other phosphonic acids or salts thereof, tripolyphosphoric acid and other phosphorus Examples thereof include an acid or a salt thereof, a polycarboxylic acid such as polyacrylic acid, polymethacrylic acid and polymaleic acid, or a salt thereof. From the viewpoint of versatility, the chelating agent (C) is preferably one or more selected from aminopolyacetic acid and a salt thereof, and more preferably one or two selected from ethylenediaminetetraacetic acid (EDTA) and a salt thereof. That is all. Examples of the counterion of these salts include alkali metals, quaternary amines, alkanolamines, etc., but alkanolamine salts are preferable from the viewpoint of corrosion resistance to medical instruments. Further, a monoethanolamine salt is preferable. These can be used alone or in combination of two or more.

In one embodiment, the content (% by mass) of the chelating agent (C) contained in the detergent for medical instruments is 0 with respect to the mass of the detergent for medical instruments from the viewpoint of the effect of removing protein stains and the cost. It is by mass% or more and 5% by mass or less, more preferably 0.005% by mass or more and 2% by mass or less, and further preferably 0.01% by mass or more and 1% by mass or less. As the content of the chelating agent (C), an acid equivalent amount is used.

As the enzyme stabilizer (D), a monosaccharide, a polysaccharide or a boron compound can be used. The monosaccharide and polysaccharide may be substituted or unsubstituted, and may be branched or linear. Examples of monosaccharides and polysaccharides include dextrin, glucose, mannose and the like. The boron compound may be substituted or unsubstituted. Examples of the boron compound include boric acid, diboron trioxide, boronic acid, and salts thereof. In one embodiment, the content (mass%) of the enzyme stabilizer (D) contained in the cleaning agent for medical instruments is 0% by mass or more 10 with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. It is preferably 0.05% by mass or more, more preferably 0.05% by mass or more and 5% by mass or less, and particularly preferably 0.1% by mass or more and 3% by mass or less.

The anticoagulant (E) includes a sugar (E-1), a nonionic surfactant (E-2), an organic acid or a salt thereof (E-3), an inorganic oxo acid or a salt thereof (E-4). , Glycerin (E-5) is included.

The sugar (E-1) includes allose, altrose, mannose, growth, idose, galactose, tarose, monosaccharides having 3 to 5 carbon atoms, ketose having 6 carbon atoms, polysaccharides having disaccharides or more, and 4 carbon atoms. Examples thereof include ~ 12 sugar alcohols. Examples of monosaccharides having 3 to 5 carbon atoms include glyceraldehyde, erythrulose, erythrose, ribose, xylulose, and xylulose. Examples of ketose having 6 carbon atoms include fructose and sorbose. Examples of the disaccharide or higher polysaccharide include sucrose, lactose, trehalose, cellobiose, sophorose, raffinose, maltotriose, carboxylmethylcellulose, starch, pullulan, pectin, glucomannan and the like. Examples of sugar alcohols having 4 to 12 carbon atoms include sorbitol, xylitol, pentaerythritol, maltitol, lactitol, sucralose and the like.

Examples of the non-surface active agent (E-2) include higher alcohol alkylene oxide adduct, alkyl (or alkenyl) phenol alkylene adduct adduct, styrated phenol alkylene oxide adduct, styrated alkyl phenol alkylene adduct adduct, and higher alkyl (or higher alkyl (or alkenyl) adduct. Higher alkenyl) amine alkylene oxide adduct, fatty acid alkylene oxide adduct, fatty acid amide alkylene oxide adduct, polypropylene glycol alkylene oxide adduct, (mono or poly) glycerol fatty acid adduct or alkylene oxide adduct thereof, sucrose fatty acid ester or its Examples thereof include an alkylene oxide adduct, a sorbitan fatty acid ester, or an alkylene oxide adduct thereof.

Here, the higher alcohol is usually a linear or branched unsaturated or saturated higher alcohol having 8 to 24 carbon atoms, and the alkyl or alkenylphenol is usually a linear or branched alkyl group or alkenyl group having 6 to 22 carbon atoms. The higher alkyl or higher alkenylamine is usually a linear or branched higher alkyl or higher alkenylamine having 8 to 24 carbon atoms, and the fatty acid is usually an unsaturated or saturated fatty acid having 8 to 24 carbon atoms. The weight average molecular weight of polypropylene glycol is 900 to 5000.

Examples of the alkyleneoxy group of the alkylene oxide adduct include ethyleneoxy group, propyleneoxy group, butyleneoxy group, and styreneoxy group, which may be used alone or in combination of two or more. When two or more kinds are used, the addition form of the alkylene oxide is not limited, and examples thereof include random addition, block addition, and a method of mixing random and block. The number of moles of the alkylene oxide adduct added is 1 to 1000.

Examples of the organic acid or a salt thereof (E-3) include amino acids, aminocarboxylic acids, keto acids, oxycarboxylic acids, polycarboxylic acids, saturated or unsaturated fatty acids having 1 to 24 carbon atoms, and salts thereof. .. Examples of the organic acid include organic compounds having an acidic group such as a carboxy group, a sulfo group, a phosphoric acid group, a thiol group, and a phenolic hydroxyl group in the molecule, and examples of these salts include alkali metal salts. Examples thereof include ammonium salts and alkanolamine salts.

More specifically, examples of the organic compound having a carboxy group include amino acids, aminocarboxylic acids, keto acids, oxycarboxylic acids, polycarboxylic acids, saturated or unsaturated fatty acids having 1 to 24 carbon atoms, and the like. Examples of the organic compound having a sulfo group include benzenesulfonic acid, linear alkylbenzenesulfonic acid, α-olefin sulfonic acid, sulfuric acid monoester and the like. Examples of the organic compound having a phosphoric acid group include adenylic acid, ethidroic acid, phosphinic acid monoester salt, phosphoric acid monoester, and phosphoric acid diester. Examples of the organic compound having a thiol group include 4-mercaptoacetophenone, thiosalicylic acid, thiobenzoic acid, thioglycolic acid and the like. Examples of the organic compound having a phenolic hydroxyl group include phenol, 2-naphthol, catechol and the like. Examples of the alkali metal salt include sodium salt and potassium salt. Examples of the alkanolamine salt include monoethanolamine salt, diethanolamine salt, triethanolamine salt and the like.

Examples of the inorganic oxoacid or a salt thereof (E-4) include phosphates, hypophosphates, pyrophosphates, polyphosphates, sulfates and the like. Examples of the inorganic oxo acid are oxo acids such as phosphorus, sulfur, nitrogen, boron, chlorine, bromine, iodine and silicon, and examples of these salts include alkali metal salts, ammonium salts and alkanolamine salts.

More specifically, examples of phosphorus oxoacids include hypophosphite, phosphite, phosphoric acid, pyrophosphoric acid, and polyphosphates having a degree of polymerization of 3 to 6. Examples of sulfur oxoacids include hyposulfuric acid, sulfite, sulfuric acid, persulfuric acid, pyrosulfuric acid, disulfurous acid, thiosulfuric acid, sulfamic acid, amidosulfuric acid, and dithionous acid. Examples of the oxo acid of nitrogen include nitrite and nitric acid. Examples of the oxo acid of boron include metaboric acid, boric acid, and perboric acid. Examples of the oxo acid of chlorine include hypochlorous acid, chloric acid, chloric acid, perchloric acid and the like. Examples of the bromic acid of bromine include hypobromous acid, bromous acid, bromic acid, and perbromic acid. Examples of the oxo acid of iodine include hypoiodous acid, iodic acid, and periodic acid. Examples of the oxo acid of silicon include ortho-silicic acid, meta-silicic acid, meta-silicic acid and the like. Examples of the alkali metal salt include sodium salt, potassium salt and the like, and examples of the alkanolamine salt include monoethanolamine salt, diethanolamine salt, triethanolamine salt and the like.

In one embodiment, the cleaning agent for medical instruments can clean blood stains more efficiently by containing the anticoagulant (E). For example, the content (mass%) of glycerin (E-5) contained in the cleaning agent for medical instruments is 0% by mass with respect to the mass of the cleaning agent from the viewpoint of the blood coagulation preventing effect and the coagulating blood dissolving effect. It is 80% by mass or less.

As the metal corrosion inhibitor (F), for example, silicate can be used. Examples of the silicate include alkali metal silicates. As the alkali metal silicate, a compound in which n of _{M 2} O · nSiO _{2 is 0.3 to 5 is used.} Further, a more preferable value of n is 1 to 3. From the viewpoint of light metal corrosion suppression performance, n is preferably 0.3 or more, and from the viewpoint of preventing scale generation derived from silicic acid, n is preferably 5 or less. Examples of the alkali metal silicate include potassium orthosilicate, sodium orthosilicate, sodium sesquisilicate, potassium sesquisilicate, sodium metasilicate, potassium metasilicate, and sodium No. 1 sodium silicate and sodium No. 2 silicate specified in JIS K1408. No. 3 sodium silicate, trade name manufactured by Nippon Kagaku Kogyo Co., Ltd.: 1K potassium silicate, 2K potassium silicate, A potassium silicate (pure content 40%) (molar ratio K ₂ O: SiO ₂ = 1: 3), etc. Be done. In one embodiment, the content (mass%) of the metal corrosion inhibitor (F) contained in the cleaning agent for medical instruments is 0% by mass or more and 10% by mass with respect to the mass of the cleaning agent from the viewpoint of corrosion suppression performance. % Or less is preferable.

The low molecular weight polyol (G) contains an alcohol compound having at least two hydroxyl groups and having 2 to 30 carbon atoms.

Examples of the low molecular weight polyol (G) include ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, propylene glycol, dipropylene glycol, tripropylene glycol, propanediol, butanediol, methylpropanediol, pentanediol, and methylbutane. Diol, ethylpropanediol, dimethylpropanediol, hexanediol, methylpentanediol, ethylmethylpropanediol, dimethylbutanediol, heptanediol, propylmethylpropanediol, isopropylmethylpropanediol, octanediol, ethylhexanediol, sec-butylbutyl Propanediol, dimethylhexanediol, trimethylpentanediol, nonanediol, octanediol, ethyl (2-methyl) propylpropanediol, trimethylhexanediol, butylethylpropanediol, diethylpentanediol, methyloctanediol, decanediol, dimethyloctanediol , Undecanediol, ethylmethyloctanediol, dodecanediol, diethyloctanediol, trimethylnonanediol, tetradecanediol, pentadecanediol, hexadecanediol, heptadecanediol, octadecanediol, eicosandiol, docosandiol, tetracosanediol, etc. Examples thereof include saturated diols and condensates thereof.

In addition, butenediol, methylenepropanediol, butinediol, hexenediol, methylpentenediol, hexadienediol, octenediol, dimethylhexenediol, decenediol, dimethyloctenediol, tetradecenediol, hydroxyoctadesenol, pentynediol, Hexindiol, methylpentindiol, heptindiol, dimethylpentindiol, dimethylhexindiol, decinediol, dimethyloctinediol, tetramethyloctinediol, tetramethyldecinediol, tetramethyldodecinediol, tetraisopropyloctinediol , Unsaturated diols such as diethyltetradecine diol and the like.

In addition, aliphatic cyclic diols such as cyclopentanediol, cyclohexanediol, cycloheptandiol, norbornandiol, cyclooctanediol, cyclodecanediol, cyclooctenediol, decalindiol, limonene glycol, terpendiol, bicyclohexanediol, and cyclododecanediol, etc. Can be mentioned.

In addition, glycerin, butanetriol, methylpropanetriol, pentantriol, methylbutanetriol, trimethylolethane, hexanetriol, ethylbutanetriol, trimethylolpropane, propylheptantriol, dimethylpentanetriol, triethanolamine, triisopropanolamine, etc. Triol alcohol and the like can be mentioned.

In addition, erythritol, pentaerythritol, pentatetrol, hexatetrol, pentantetrol, hexanetetrol, diglycerin, sorbitan, N, N, N', N'-tetrakis (2-hydroxypropyl) ethylenediamine, N, N , N', N'-tetrakis (hydroxyethyl) ethylenediamine and other tetrahydric alcohols and the like.

Examples thereof include pentahydric alcohols such as adonitol, arabitol, xylitol and triglycerin, and hexahydric alcohols such as dipentaerythritol, sorbitol, mannitol, iditol, inositol, darcitol, tarose and allose.

In addition, methyl glyceryl ether, ethyl glyceryl ether, propyl glyceryl ether, isopropyl glyceryl ether, butyl glyceryl ether, isobutyl glyceryl ether, pentyl glyceryl ether, hexyl glyceryl ether, heptyl glyceryl ether, octyl glyceryl ether, (2-ethylhexyl) glyceryl ether, Nonyl glyceryl ether, decyl glyceryl ether, undecyl glyceryl ether, dodecyl glyceryl ether, tridecyl glyceryl ether, tetradecyl glyceryl ether, hexadecyl glyceryl ether, octadecyl glyceryl ether, eicosyl glyceryl ether, allyl glyceryl ether, undecyl glyceryl ether , Oleyl glyceryl ether, cyclohexyl glyceryl ether, phenyl glyceryl ether and other glycerin monoethers and the like.

Further, N-substituted diethanolamines such as N-methyldiethanolamine, N-ethyldiethanolamine, N-propyldiethanolamine, N-isopropyldiethanolamine, N-butyldiethanolamine, N-cyclohexyldiethanolamine, N- (2-ethylhexyl) diethanolamine and the like can be mentioned. Be done.

In addition, N-methyldiisopropanolamine, N-ethyldiisopropanolamine, N-propyldiisopropanolamine, N-isopropyldiisopropanolamine, N-butyldiisopropanolamine, N-cyclohexyldiisopropanolamine, N- (2-ethylhexyl) ) N-substituted diisopropanolamines such as diisopropanolamine and the like can be mentioned.

Further, N, N-di-substituted aminopropanediols such as dimethylaminopropanediol, diethylaminopropanediol, dipropylaminopropanediol, diisopropylaminopropanediol and dibutylaminopropanediol can be mentioned. These exemplified low molecular weight polyols (G) may also include position isomer compounds.

In one embodiment, the content (mass%) of the low molecular weight polyol (G) contained in the cleaning agent for medical instruments is 0% by mass or more and 80% by mass with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. It is preferably mass% or less.

Cleaning aids (builders) (H) include polycarboxylic acids (acrylic acid homopolymers, maleate homopolymers, etc.), polyvalent carboxylates (citric acid, malic acid, etc.), and alkaline builders (caustic soda). , Soda ash, ammonia, triethanolamine, sodium tripolyphosphate, sodium silicate, etc.) and the like. In one embodiment, the content (% by mass) of the builder (H) contained in the cleaning agent is preferably 0% by mass or more and 20% by mass or less with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. ..

Examples of the defoaming agent (I) include silicone-based defoaming agents, polyoxyalkylene-based defoaming agents, and mineral oil-based defoaming agents. In one embodiment, the content (mass%) of the defoaming agent (I) contained in the cleaning agent for medical instruments is 0% by mass or more 10 with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. It is preferably mass% or less.

Examples of the pH adjuster (J) include sulfuric acid, hydrochloric acid, citric acid, lactic acid, pyruvic acid, formic acid, sodium chloride, potassium chloride, monoethanolamine and diethanolamine. In one embodiment, the content (mass%) of the pH adjuster (J) contained in the cleaning agent for medical instruments is 0% by mass or more 25 with respect to the mass of the cleaning agent for medical instruments from the viewpoint of detergency. It is preferably 0% by mass or more, more preferably 0% by mass or more and 15% by mass or less, and particularly preferably 0% by mass or more and 10% by mass or less.

The concentration of the fluoroalcohol when acting on the protein aggregate may be 0.1% by mass or more, preferably 1% by mass or more, and further preferably 8% by mass or more. The concentration of the fluoroalcohol when acting on the protein aggregate is more preferably 29% by mass or more. The time for the fluoroalcohol to act on the protein aggregate is not particularly limited. The time for the fluoroalcohol to act on the protein aggregate is effective even for a short time. For example, the time for allowing the fluoroalcohol to act on the protein aggregate may be 30 seconds or longer, preferably 1 minute or longer, more preferably 20 minutes or longer, and particularly preferably 30 minutes or longer.

In one embodiment, the cleaning agent for medical instruments may contain an active ingredient of a commercially available disinfectant as an additive. Examples of the active ingredient of a commercially available disinfectant include an active ingredient of a high-level disinfectant (for example, glutarualdehyde, orthophthalaldehyde, peracetic acid or peracetic acid), and an active ingredient of a medium-level disinfectant (hypochlorite). Sodium, ethanol, povidone iodine), active ingredients of low-level disinfectants (quaternary ammonium, chlorhexidine gluconate). In one embodiment, the content (mass%) of these active ingredients contained in the cleaning agent for medical instruments is 0% by mass or more and 99.9% by mass or less with respect to the mass of the cleaning agent for medical instruments. It is also preferable, 0% by mass or more and 99% by mass or less, 0% by mass or more and 92% by mass or less is particularly preferable, and 0% by mass or more and 71% by mass or less is further preferable.

In one embodiment, the fluoroalcohol contained in the cleaning agent for medical instruments, especially HFIP, is effective for cleaning medical instruments contaminated with protein aggregates or to which protein aggregates are attached. In addition, HFIP can lyse organic substances such as proteins and cell tissues. Therefore, it becomes easy to remove organic substances such as proteins and cell tissues adhering to the object to be cleaned, and the cleaning power can be improved. HFIP is a stable low molecular weight compound having a molecular weight of 168 and has high storage stability. Further, since it has good thermal stability, the cleaning temperature is not limited, and the cleaning power can be further improved. HFIP is less corrosive to metals and has less effect on the material of the object to be cleaned. Furthermore, HFIP is nonflammable and easy to manage safety in use.

[Washing method]
The cleaning agent for medical devices according to the present embodiment is used for cleaning various medical devices (including, for example, an endoscope), as well as for cleaning objects including animal medical devices, meat processing tools, and cooking tools. Can be used. However, the present invention is not limited to this, and can be widely applied as a countermeasure against infection of protein aggregates. For example, it can be used for cleaning an operating room in the medical field, linen such as beds and sheets, and cleaning an object including disinfection of human hands.

In one embodiment, the cleaning agent for medical instruments may be, for example, manual cleaning (including immersion cleaning), ultrasonic cleaning, jet cleaning, shower cleaning, steam cleaning, vacuum cleaning, degassing cleaning, washer-disinfector, or It can be used in any of these two or more combinations of cleaning methods, but is not limited thereto. For example, a cleaning method using a washer-disinfector generally consists of steps of pre-cleaning, main cleaning, rinsing, and disinfection. The cleaning agent for medical instruments according to the present embodiment can be used for pre-cleaning and main cleaning, and can also be used in combination with other cleaning agents for pre-cleaning or main cleaning. Here, the permissible concentration of the fluorine-based alcohol contained in the cleaning agent for medical instruments is the same as the concentration at which the protein aggregate can be altered, and specifically, it may be 0.1% by mass or more. 1, 1% by mass or more is preferable, 8% by mass or more is more preferable, and 29% by mass or more is most preferable.

In one embodiment, the cleaning agent for medical instruments is used in the pre-cleaning step and / or the main cleaning step of the cleaning method to efficiently obtain biological blood or body fluid, fat, prion protein and infectious amyloid from the object. It is possible to remove proteins such as proteins, organic substances such as cell tissues, microorganisms, viruses, and the like.

However, the cleaning agent for medical instruments according to the present embodiment is not limited to the above-mentioned uses, and may be used in the same manner as known antiviral agents, antibacterial agents, bactericidal agents, disinfectants, fungicides, etc., depending on the object to be cleaned. can do. For example, a method of spraying on an object to be cleaned, a method of applying, a method of impregnating the object, a method of immersing the object, a method of exposing the object to high-pressure steam, and the like, which are usually adopted, can be used as they are. ..

The temperature at which the fluorine-based alcohol contained in the cleaning agent for medical instruments alters the protein aggregate is not particularly limited, and a temperature of room temperature or higher (for example, 20 ° C or higher) is preferable. The higher the temperature at which the fluoroalcohol and the protein aggregate are brought into contact with each other, the more easily the protein aggregate is altered. Further, the temperature may be higher than the boiling point of the fluoroalcohol, that is, vaporized into vapor and brought into contact with the protein aggregate. In addition, the optimum temperature can be selected depending on the solvent and additives contained in the cleaning agent for medical instruments.

[Washing device]
In one embodiment, it is possible to provide a cleaning device including the action of the above-mentioned cleaning agent for medical instruments according to the present embodiment.

FIG. 1 is a schematic view showing a cleaning device 1 according to an embodiment of the present invention. For example, the cleaning device 1 may be for cleaning a medical device. As shown in FIG. 1, the cleaning device 1 includes a cleaning tank 10 having a storage unit 20 for accommodating an object (medical device), and a cleaning agent supply device 40 for supplying a cleaning agent for medical devices into the cleaning tank 10. , Equipped with. The washing tank 10 has a water storage unit 12 that stores wash water under the storage unit 20, and a water supply source and water supply as a washing water supply means for supplying water and hot water sent from the hot water supply source to the water storage unit 12 as wash water. The pipe 14, the hot water supply pipe 16, the cleaning nozzle 22 that injects the cleaning water of the water storage unit 12 onto the object housed in the cleaning tank 10, the cleaning pump 18 that sends the cleaning water of the water storage unit 12 to the cleaning nozzle 22. To be equipped with.

In the cleaning method using the cleaning device 1, the medical equipment is cleaned by the preliminary cleaning step, the main cleaning step, and the rinsing cleaning step, and then disinfected with boiling water by the disinfection step. In one embodiment, the cleaning agent for medical instruments can be used for pre-cleaning and main cleaning, and can also be used for pre-cleaning or main cleaning in combination with other cleaning agents. In addition, when the cleaning agent for medical instruments contains a plurality of components, some of the components or each component can be used for pre-cleaning or main cleaning from separate lines.

When the cleaning process is started, the cleaning device 1 first executes the preliminary cleaning process. The cleaning device 1 opens the water supply valve and starts the water supply treatment in the preliminary cleaning process. As a result, the water from the water supply source is sent out to the water storage unit 12 of the washing tank 10 through the water supply pipe 14. The cleaning device 1 determines whether or not the water in the water storage unit 12 has reached a predetermined water level by detecting the float switch. When the cleaning device 1 determines that the water in the water storage unit 12 has reached a predetermined water level, the cleaning device 1 closes the water supply valve and ends the water supply process. The water in the pre-cleaning step may be at room temperature (eg 20 ° C.).

As a preliminary cleaning process, the cleaning device 1 supplies the cleaning agent into the cleaning tank 10 by the cleaning agent supply device 40, and operates the cleaning pump 18 for a predetermined time. As a result, the cleaning water in the water storage unit 12 is sent to the cleaning nozzle 22 via the circulation pipe, and the cleaning nozzle 22 injects the cleaning water onto the medical device while rotating to perform cleaning. The cleaning water containing the cleaning agent in the cleaning tank 10 falls and returns to the water storage unit 12. After the pre-cleaning treatment, the cleaning device 1 operates the drainage pump for a predetermined time as the wastewater treatment. As a result, the washing water in the water storage unit 12 is discharged to the outside of the machine through the drain pipe 34.

When the pre-cleaning process is completed, the cleaning device 1 executes the main cleaning process. In the main cleaning step, the cleaning device 1 opens the water supply valve and the hot water supply valve to start the water supply process in the main cleaning step. As a result, the water from the water supply source and the hot water from the hot water supply source are sent out to the water storage unit 12 of the washing tank 10 through the water supply pipe 14 and the hot water supply pipe 16. The cleaning device 1 determines whether or not the water in the water storage unit 12 has reached a predetermined water level by detecting the float switch. When the cleaning device 1 determines that the water in the water storage unit 12 has reached a predetermined water level, the cleaning device 1 closes the water supply valve and the hot water supply valve to end the water supply process. The water in this washing step is preferably 25 ° C. or higher.

As the main cleaning process, the cleaning device 1 supplies the cleaning agent into the cleaning tank 10 by the cleaning agent supply device 40, and operates the cleaning pump 18 for a predetermined time. As a result, the cleaning water in the water storage unit 12 is sent to the cleaning nozzle 22 via the circulation pipe, and the cleaning nozzle 22 injects the cleaning water onto the medical device while rotating to perform cleaning. The cleaning water containing the cleaning agent in the cleaning tank 10 falls and returns to the water storage unit 12. After the main cleaning treatment, the cleaning device 1 operates the drainage pump for a predetermined time as the wastewater treatment. As a result, the washing water in the water storage unit 12 is discharged to the outside of the machine through the drain pipe 34.

This cleaning step may be executed multiple times. In this case, for example, the main cleaning treatment may be performed a plurality of times by using different cleaning agents. When the cleaning agent for medical instruments according to the present embodiment is used in the first main cleaning step, the main cleaning treatment may be performed with cleaning water at 60 ° C. or higher. When an enzyme-based cleaning agent is used in the second main cleaning step, the main cleaning treatment may be performed with cleaning water at about 30 to 40 ° C. (for example, 37 ° C.).

When the main cleaning process is completed, the cleaning device 1 executes the rinsing process. In the rinsing washing step, the washing device 1 opens the hot water supply valve and starts the hot water supply process in the rinsing step. As a result, the hot water of the hot water supply source is sent to the water storage unit 12 of the washing tank 10 through the hot water supply pipe 16. The cleaning device 1 determines whether or not the hot water in the water storage unit 12 has reached a predetermined water level by detecting the float switch. When the cleaning device 1 determines that the hot water in the water storage unit 12 has reached a predetermined water level, the cleaning device 1 closes the hot water supply valve and ends the hot water supply process.

The cleaning device 1 operates the cleaning pump 18 for a predetermined time as a rinsing process. As a result, the hot water in the water storage unit 12 is sent to the cleaning nozzle 22 via the circulation pipe, and the cleaning nozzle 22 injects hot water onto the medical device while rotating to wash away the cleaning water containing the cleaning agent. After the rinsing treatment, the cleaning device 1 operates the drainage pump for a predetermined time as the wastewater treatment. As a result, the rinse water in the water storage unit 12 is discharged to the outside of the machine through the drain pipe 34.

When the rinsing process is completed, the cleaning device 1 executes the disinfection process. In the disinfection step, the cleaning device 1 opens the hot water supply valve and starts the hot water supply process in the disinfection step. As a result, the hot water of the hot water supply source is sent to the water storage unit 12 of the washing tank 10 through the hot water supply pipe 16. The cleaning device 1 determines whether or not the hot water in the water storage unit 12 has reached a predetermined water level by detecting the float switch. When the cleaning device 1 determines that the hot water in the water storage unit 12 has reached a predetermined water level, the cleaning device 1 closes the hot water supply valve and ends the hot water supply process. The boiling water in the disinfection step is preferably 60 ° C. or higher.

The cleaning device 1 operates the cleaning pump 18 for a predetermined time as a disinfection process. As a result, the hot water in the water storage unit 12 is sent to the cleaning nozzle 22 via the circulation pipe, and the cleaning nozzle 22 injects the hot water into the medical device while rotating. After the disinfection treatment, the cleaning device 1 operates the drainage pump for a predetermined time as the wastewater treatment. As a result, the hot water in the water storage unit 12 is discharged to the outside of the machine through the drain pipe 34.

The cleaning device 1 efficiently removes protein aggregates from an object by supplying the cleaning agent for medical equipment according to the present embodiment into the cleaning tank 10 by the cleaning agent supply device 40, and at the same time, living body-derived blood and blood and the like. It can remove proteins such as body fluids, fats, prion proteins and infectious amyloids, organic substances such as cell tissues, microorganisms, viruses and the like. The cleaning device 1 can also use a cleaning agent for medical instruments according to the present embodiment and a known cleaning agent in combination.

The cleaning device 1 can also be used for cleaning animal medical utensils, meat processing utensils, and cooking utensils.

[Soil cleaning agent]
The molecular structure altering agent according to the present invention described above can be used as a soil cleaning agent in one embodiment. For example, it has been pointed out that ^{PrP Sc} may flow out into the environment from the carcasses of animals that have developed BSE, sheep scrapie, CWD, etc., and pollute the soil. At this time, no drug is known that can effectively remove protein aggregates from soil. In one embodiment, the soil cleaning agent may also include common additives described as known techniques. These also include pesticides, fertilizers, fungicides, disinfectants, etc. used in soil.

In one embodiment, the soil cleaning agent contains the above-mentioned molecular structure altering agent according to the present invention, and may further contain a solvent. The solvent can be selected from substances that can dilute the molecular structure altering agent, and examples thereof include the same solvents as those mentioned in the above-mentioned cleaning agents for medical devices. These may be one or more, but are not limited thereto.

In one embodiment, the content (% by mass) of the solvent contained in the soil cleaning agent may be 0% by mass or more and 99.9% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of cleaning property. Often, 0% by mass or more and 99% by mass or less is preferable, 0% by mass or more and 92% by mass or less is particularly preferable, and 0% by mass or more and 71% by mass or less is further preferable.

In one embodiment, the soil cleaning agent may contain additives in addition to the molecular structure altering agent and the solvent. Additives that can be added to soil cleaning agents include surfactants, enzymes, enzyme stabilizers, metal corrosion inhibitors, low molecular weight polyols, cleaning aids (builders), defoamers, pH adjusters, fragrances, and colorants. , Antioxidants, preservatives, bleaching agents, bleaching activators, corrosion inhibitors, dispersants, thickeners, viscosity regulators, etc., but are not limited thereto. The soil cleaning agent may contain one or more additives. In one embodiment, the soil cleaning agent can further improve the cleaning power by containing a molecular structure altering agent, a solvent, and the above-mentioned additives.

Examples of the surfactant (A) include the same surfactants (A) as those mentioned in the above-mentioned cleaning agents for medical devices. As the surfactant (A), one kind or two or more kinds can be used.

The content (mass%) of the surfactant (A) contained in the soil cleaning agent is preferably 0% by mass or more and 10% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of detergency, and more preferably. It is 0.1% by mass or more and 5% by mass or less.

Examples of the enzyme (B) include the same enzymes (B) as those mentioned in the above-mentioned cleaning agents for medical devices. As the enzyme (B), one type or two or more types can be used.

In one embodiment, the content (mass%) of the enzyme (B) contained in the soil cleaning agent is preferably 0% by mass or more and 10% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of cleanability. More preferably, it is 0.05% by mass or more and 5% by mass or less, and particularly preferably 0.1% by mass or more and 3% by mass or less.

Examples of the enzyme stabilizer (C) include the same enzyme stabilizer (D) mentioned in the above-mentioned cleaning agent for medical devices. As the enzyme stabilizer (C), one kind or two or more kinds can be used. In one embodiment, the content (mass%) of the enzyme stabilizer (C) contained in the soil cleaning agent is 0% by mass or more and 10% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of cleanability. It is preferable, more preferably 0.05% by mass or more and 5% by mass or less, and particularly preferably 0.1% by mass or more and 3% by mass or less.

Examples of the metal corrosion inhibitor (D) include the same as the metal corrosion inhibitor (F) mentioned in the above-mentioned cleaning agent for medical devices. As the metal corrosion inhibitor (D), one type or two or more types can be used. In one embodiment, the content (mass%) of the metal corrosion inhibitor (F) contained in the soil cleaning agent is 0% by mass or more and 10% by mass with respect to the mass of the soil cleaning agent from the viewpoint of corrosion suppression performance. The following is preferable.

Examples of the low-molecular-weight polyol (E) include those similar to the low-molecular-weight polyol (G) mentioned in the above-mentioned cleaning agent for medical devices. As the low molecular weight polyol (E), one kind or two or more kinds can be used.

In one embodiment, the content (mass%) of the low molecular weight polyol (E) contained in the soil cleaning agent is 0% by mass or more and 80% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of detergency. preferable.

Examples of the cleaning aid (builder) (F) include the same cleaning aid (builder) (H) mentioned in the above-mentioned cleaning agent for medical devices. As the builder (F), one type or two or more types can be used. In one embodiment, the content (mass%) of the builder (F) contained in the soil cleaning agent is preferably 0% by mass or more and 20% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of detergency.

Examples of the defoaming agent (G) include those similar to the defoaming agent (I) mentioned in the above-mentioned cleaning agent for medical devices. As the defoaming agent (G), one kind or two or more kinds can be used. In one embodiment, the content (mass%) of the defoaming agent (G) contained in the soil cleaning agent is 0% by mass or more and 10% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of detergency. preferable.

Examples of the pH adjuster (H) include those similar to the pH adjuster (J) mentioned in the above-mentioned cleaning agent for medical devices. As the pH adjuster (H), one type or two or more types can be used. In one embodiment, the content (mass%) of the pH adjuster (H) contained in the soil cleaning agent is 0% by mass or more and 25% by mass or less with respect to the mass of the soil cleaning agent from the viewpoint of cleanability. It is preferable, more preferably 0% by mass or more and 15% by mass or less, and particularly preferably 0% by mass or more and 10% by mass or less.

The concentration of the fluoroalcohol when acting on the protein aggregate may be 0.1% by mass or more, preferably 1% by mass or more, and further preferably 8% by mass or more. The concentration of the fluoroalcohol when acting on the protein aggregate is more preferably 29% by mass or more. The time for the fluoroalcohol to act on the protein aggregate is not particularly limited. The time for the fluoroalcohol to act on the protein aggregate is effective even for a short time. For example, the time for allowing the fluoroalcohol to act on the protein aggregate may be 30 seconds or longer, preferably 1 minute or longer, more preferably 20 minutes or longer, and particularly preferably 30 minutes or longer. In order to increase the effect, it is also preferable to use a plurality of times in stages with intervals of use on a daily or weekly basis or even on a seasonal basis.

In one embodiment, the soil cleaning agent may contain the active ingredient of a commercially available disinfectant as an additive. Examples of the active ingredient of the commercially available disinfectant include the same active ingredients of the commercially available disinfectant mentioned in the above-mentioned cleaning agents for medical devices. In one embodiment, the content (% by mass) of these active ingredients contained in the soil cleaning agent may be 0% by mass or more and 99.9% by mass or less with respect to the mass of the soil cleaning agent, and is 0. It is preferably 0% by mass or more and 99% by mass or less, particularly preferably 0% by mass or more and 92% by mass or less, and further preferably 0% by mass or more and 71% by mass or less.

In one embodiment, the fluoroalcohol contained in the soil cleaning agent, especially HFIP, is effective for cleaning soil contaminated with protein aggregates. HFIP is a stable low molecular weight compound having a molecular weight of 168 and has high storage stability. Further, since it has good thermal stability, the cleaning temperature is not limited, and the cleaning power can be further improved. Furthermore, HFIP is nonflammable and easy to manage safety in use.

[Washing method]
The soil cleaning agent according to the present embodiment can be used to clean soil contaminated with protein aggregates or soil that may be contaminated. Alternatively, the soil may be washed prophylactically with the soil cleaning agent according to the present embodiment. In one embodiment, the soil cleaning agent may be diluted with the above solvent and used for cleaning. In one embodiment, the soil cleaning agent or solvent and the soil cleaning agent according to the present embodiment are added to the soil to be cleaned, mixed, and the washed soil is separated and dried to alter the protein agglomerates. The soil can be washed.

In one embodiment, at the time of washing, the concentration of the fluorinated alcohol or the compound represented by the general formula (1) may be 0.01% by mass or more, preferably 0.1% by mass or more, and 1% by mass or more. Is particularly preferable. It can be adjusted arbitrarily while considering the degree of soil contamination at the time of use, the time of contamination, the surrounding environment of the soil, and the like.

In one embodiment, the fluoroalcohol or the compound represented by the general formula (1) can also be used as a soil fumigant. For example, the soil can be washed by fumigating the soil suspected of being contaminated with a fluorine-based alcohol or a soil fumigant containing a compound represented by the general formula (1).

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples as long as the gist of the present invention is not exceeded.

<Molecular structure altering agent>
HFIP (Central Glass Co., Ltd., purity 99% or higher) was used as a compound that alters the molecular structure of protein aggregates.

<Cells derived from mouse neuroblastoma>
In cell cultures from murine neuroblastoma, and Neuro 2a (N2a) cells producing normal prion protein (PrP ^C), the ScNeuro 2a (ScN2a) cells that persist producing an abnormal prion protein ^{(PrP Sc)} 2 A type of cell line (ATCC) was prepared. The two cell lines were humidified in Eagle's Minimal Essential Medium (E-MEM) (Fuji Film Wako Pure Chemical Industries, Ltd.) containing 10% FBS (fetal bovine serum) and 100 Units / ml Penicillin, 100 μg / ml Streptomycin. The cells were cultured under 37 ° C. and 5% CO _{2 conditions.} A cell culture dish (AGC Techno Glass Co., Ltd.) and a cell culture microplate (6 wells or 24 wells, AGC Techno Glass Co., Ltd.) were used as cell culture vessels.

^{1 × 10 5} cells per well were seeded in a 24-well plate for cell culture, and N2a cells and ScN2a cells were incubated with E-MEM under the conditions of 37 ° C. in _{5% CO 2 for 2 days.}

[Example 1]
In the evaluation of susceptibility to HFIP described above, various media having final concentrations of HFIP in the medium of 0 mM, 10 mM, 20 mM, 25 mM, 30 mM, and 40 mM were used. Here, "M" indicates a molar concentration (mol / L). These were added to ScN2a cells and cell culture was carried out for 24 hours.

[Comparative Example 1]
The procedure was the same as in Example 1 except that the cells used were changed to N2a cells.

<Evaluation of susceptibility to HFIP>
The susceptibility of each cell to HFIP was evaluated by cell staining with crystal violet. Here, crystal violet is a reagent that binds to the cell membrane of living cells and dyes purple. After washing once with PBS (phosphate buffered saline), 100% methanol was added and the cells were fixed by incubating at room temperature for 15 minutes. Next, methanol was removed, a 0.05% (weight / volume%) crystal violet solution (Fujifilm Wako Pure Chemical Industries, Ltd.) was added, and the cells were allowed to stand at room temperature for 15 minutes to stain the cells. After removing the crystal violet solution with a pipette, the number of viable cells was evaluated from the stained area for each well. A photograph of the appearance of the plate of Example 1 after cell staining is shown in the lower part of FIG. 2, and a photograph of the appearance of the plate of Comparative Example 1 is shown in the upper part of FIG.

As shown in FIG. 2, the upper limit of the concentration of HFIP in which N2a cells can survive was 25 mM, while the upper limit of the concentration of HFIP in which ScN2a cells could survive was 20 mM. This result indicates that N2a cells and ScN2a cells have different sensitivities to HFIP. This difference in susceptibility to HFIP indicates that HFIP can be used to detect abnormally structured prion proteins. Therefore, it has been shown that it can be suitably used for simple screening and diagnosis of protein misfolding diseases having a β-sheet structure or an amyloid structure.

<Evaluation of protease resistance>
^{3 × 10 5} cells per well were seeded in a 6-well plate for cell culture, and N2a cells and ScN2a cells were incubated with E-MEM at 37 ° C. and 5% CO _{2 for} 2 days.

[Example 2]
The medium of ScN2a cells was replaced with a medium having a final concentration of HFIP of 6 mM and incubated for 24 hours.

[Comparative Example 2]
The procedure was the same as in Example 2 except that the cells used were changed to N2a cells.

After incubation for 24 hours, the medium was removed by suction, and each cell was washed with cooled PBS. Next, 500 μL of cooled RIPA buffer (Fujifilm Wako Pure Chemical Industries, Ltd.) was added per well, and the cells were lysed by allowing to stand on ice for 1 minute. The entire amount of cytolysis was collected in a 1.5 ml tube and centrifuged at 4 ° C., 3000 rpm for 10 minutes. 200 μL of the centrifugal supernatant was transferred to a new tube, proteinase K (Protease K, Nacalai Tesque, Inc.) was added to a final concentration of 10 μg / mL, and the mixture was reacted in a constant temperature bath at 37 ° C. for 30 minutes. Next, Proteinase inhibitor (Nacalai Tesque Co., Ltd.) was added, and the protease K activity was inactivated by reacting at room temperature for 5 minutes using a tube rotor.

Then, the mixture was centrifuged at 4 ° C. and 13000 rpm for 20 minutes, and the supernatant was removed with a pipette. Dissolution buffer for gel electrophoresis (150 mM Tris-HCl (Tris hydrochloride buffer) pH 6.8, 6% SDS (sodium dodecyl sulfate), 30% glycerol, 0.01% BPB (bromophenol blue)) with respect to the precipitated fraction. , 100 mM DTT (dithiotreitol)) was added, incubated at 95 ° C. for 5 minutes, and subjected to SDS-PAGE (polyacrylamide gel electrophoresis).

Western blotting was used to detect the proteinase K-resistant prion protein. A 15% acrylamide gel (DRC Co., Ltd.) was used as a sample, and the protein was run on a Tris-Glycine SDS Running Buffer (25 mM Tris, 192 mM Glycine, 0.1% SDS) at 100 V for 15 minutes to allow the protein to have a molecular weight. Separated according to.

The gel after electrophoresis was transferred to a PVDF membrane (polyvinylidene fluoride). As the PVDF film, iBlot2 Transfer Stacks, PVDF, and regular size (Cat.IB24001) were used. For transcription, iBlot Gel Transfer Device (Thermo Fisher Scientific) was used, and energization was carried out at 20 V for 1 minute, 23 V for 4 minutes, and 25 V for 2 minutes for a total of 7 minutes. The PVDF membrane after transfer was treated with PBS-Tween 20 (PBS-T) containing 5% (weight / volume%) skim milk for 30 minutes at room temperature (blocking), and anti-PrP diluted 1000-fold with PBS-T. The antibody (Cat. A03207, SPI-Bio) was added to the PVDF membrane and incubated overnight at room temperature. After washing the PVDF membrane with PBS-T with shaking once in 15 minutes and 3 times in 5 minutes, HRP-labeled anti-mouse IgG (Cat. W4021, Promega) diluted 5000-fold with PBS-T was added. Then, the mixture was incubated at room temperature for 1 hour with shaking. Then, the PVDF membrane was washed with PBS-T while shaking once in 15 minutes and 3 times in 5 minutes, and then a chemiluminescent reagent (Immobilon western chemiluminescent HRP substrate, Mark Millipore) was added to the PVDF membrane for 1 minute. After incubation, proteinase K-resistant prion protein was detected by an image analyzer (ChemiDoc Touch, Bio-Rad Laboratories, Hercules).

The results of Western blotting of Example 2 are shown in # 5 to # 8 of FIG. 3, and the results of Western blotting of Comparative Example 2 are shown in # 1 to # 4 of FIG.

As shown in FIG. 3, it was shown that the protein extracted from the N2a cells of Comparative Example 2 was degraded by proteinase K regardless of the addition of HFIP (# 1 to # 4). However, in the protein extracted from ScN2a cells of Example 2, changes were observed in the band after the action on proteinase K with or without the addition of HFIP. This indicates that the resistance to proteinase K was reduced in ScN2a cells supplemented with HFIP. Therefore, it was suggested that HFIP has an activity of alleviating the abnormal structure ^{of PrP Sc} in ScN2a cells.

The molecular structure altering agent for protein aggregates in the present invention is usefully used for diagnostic purposes of protein misfolding diseases.

1 cleaning device, 10 cleaning tank, 12 water storage section, 14 water supply pipe, 16 hot water supply pipe, 18 cleaning pump, 20 storage section, 22 cleaning nozzle, 34 drain pipe, 40 cleaning agent supply device, 100 inspection device, 110 input section, 120 detection unit, 130 storage unit, 140 display unit, 150 control unit, 160 calculation unit, 170 power supply device, 180 communication unit, 200 network, 300 server, 400 terminal

Claims (27)

1. A molecular structure altering agent containing a fluoroalcohol or a compound represented by the following general formula (1) as a compound for detecting protein aggregates.
   

   

   
   (In the general formula (1), a is 0 or 1,
   When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
   When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
   R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
   R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
   Satisfy l + s <6. )
2. The fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH.
   Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms.
   The molecular structure altering agent according to claim 1, wherein Rf and Rf'are different from each other or are the same.
3. The molecular structure altering agent according to claim 1, wherein the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.
4. The molecular structure altering agent according to claim 1, wherein the concentration of the fluorinated alcohol or the compound represented by the general formula (1) when acting on the protein aggregate is in the range of 10 pM to 100 mM.
5. The molecular structure altering agent according to claim 1, which is used for diagnosing protein misfolding disease.
6. Protein misfolding disease is sporadic Creutzfeldt-Jakob disease in humans, hereditary Gerstmann-Stroisler-Scheinker syndrome, lethal familial insomnia, iatrogenic or dietary variant CJD, bovine spongy Neurodegeneration in the spectrum of encephalopathy, sheep scrapy, deer chronic wasting disease, prion disease in animals that can develop prion disease, Alzheimer's disease, Parkinson's disease, Levy body dementia, frontotemporal lobar degeneration (FTLD) The molecular structural alteration according to claim 1, which is one disease selected from the group consisting of a disease group, muscle atrophic lateral sclerosis, Huntington chorea, polyglutamine disease, cataract, age-related yellow spot and systemic amyloidosis. Agent.
7. A method for detecting protein aggregates, which comprises allowing a fluoroalcohol or a compound represented by the following general formula (1) to act on the protein aggregates.
   

   

   
   (In the general formula (1), a is 0 or 1,
   When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
   When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
   R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
   R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
   Satisfy l + s <6. )
8. Claim that the fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH, Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms, and Rf and Rf'are different from each other or are the same. Item 7. The method for detecting protein aggregates according to Item 7.
9. The method for detecting protein aggregates according to claim 7, wherein the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.
10. The method for detecting protein aggregates according to claim 7, wherein the concentration of the fluoroalcohol or the compound represented by the general formula (1) when acting on the protein aggregates is in the range of 10 pM to 100 mM.
11. The method for detecting protein aggregates according to claim 7, which is used for diagnosing protein misfolding disease.
12. Protein misfolding disease is sporadic Creutzfeldt-Jakob disease in humans, hereditary Gerstmann-Stroisler-Scheinker syndrome, lethal familial insomnia, iatrogenic or dietary variant CJD, bovine spongy Neurodegeneration in the spectrum of encephalopathy, sheep scrapy, deer chronic wasting disease, prion disease in animals that can develop prion disease, Alzheimer's disease, Parkinson's disease, Levy body dementia, frontotemporal lobar degeneration (FTLD) Detection of protein aggregates according to claim 7, which is used for diagnosing any one or more of disease groups, muscle atrophic lateral sclerosis, Huntington chorea, polyglutamine disease, cataracts, age-related yellow spots and systemic amyloidosis. Method.
13. A test kit for detecting protein aggregates containing a fluoroalcohol or a compound represented by the following general formula (1) as a compound for detecting protein aggregates.
    

    

    
    (In the general formula (1), a is 0 or 1,
    When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
    When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
    R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
    R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
    Satisfy l + s <6. )
14. The fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH.
    Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms.
    The test kit according to claim 13, wherein Rf and Rf'are different from each other or are the same.
15. The test kit according to claim 13, wherein the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.
16. The test kit according to claim 13, wherein the concentration of the fluorinated alcohol or the compound represented by the general formula (1) when acting on the protein aggregate is in the range of 10 pM to 100 mM.
17. An inspection device for detecting protein aggregates, which comprises a detection unit containing a fluoroalcohol or a compound represented by the following general formula (1) as a compound for detecting protein aggregates.
    

    

    
    (In the general formula (1), a is 0 or 1,
    When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
    When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
    R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
    R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
    Satisfy l + s <6. )
18. The fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH.
    Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms.
    The inspection device according to claim 17, wherein Rf and Rf'are different from each other or are the same.
19. The inspection device according to claim 17, wherein the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.
20. The inspection apparatus according to claim 17, wherein the concentration of the fluorinated alcohol or the compound represented by the general formula (1) when acting on the protein aggregate is in the range of 10 pM to 100 mM.
21. A cleaning agent for medical instruments containing a fluoroalcohol or a compound represented by the following general formula (1).
    

    

    
    (In the general formula (1), a is 0 or 1,
    When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
    When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
    R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
    R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
    Satisfy l + s <6. )
22. Claims that the fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH, Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms, and Rf and Rf'are different from each other or are the same. Item 21. The cleaning agent for medical instruments according to Item 21.
23. The cleaning agent for medical devices according to claim 21, wherein the fluorine-based alcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.
24. A soil cleaning agent containing a fluorinated alcohol or a compound represented by the following general formula (1).
    

    

    
    (In the general formula (1), a is 0 or 1,
    When a = 1, R ¹ is a hydrogen atom and R ² is a hydroxyl group, or R ¹ is a hydroxyl group and R ² is a hydrogen atom.
    When a = 0, R ² is an oxygen atom that forms a double bond with the carbon atom.
    R ³ is CH _l Cl _m F _n , l is an integer from 0 to 3, m and n are integers from 1 to 3, satisfying l + m + n = 3.
    R ⁴ is CH _s Cl _t F _u , s is an integer from 0 to 3, t and u are integers from 1 to 3, satisfying s + t + u = 3.
    Satisfy l + s <6. )
25. Claim that the fluoroalcohol is represented by the general formula RfCH ₂ OH or RfRf'CHOH, Rf and Rf'represent a perfluoroalkyl group having 1 to 10 carbon atoms, and Rf and Rf'are different from each other or are the same. Item 24. The soil cleaning agent.
26. The soil cleaning agent according to claim 24, wherein the fluoroalcohol is 1,1,1,3,3,3-hexafluoro-2-propanol.
27. A method for cleaning soil using the soil cleaning agent according to claim 24.

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